Benzoquinoline inhibitors of vesicular monoamine transporter 2

ABSTRACT

The present invention relates to new benzoquinoline inhibitors of vesicular monoamine transporter 2 (VMAT2), pharmaceutical compositions thereof, and methods of use thereof.

This application is a continuation of U.S. patent application Ser. No.15/041,571, filed Feb. 11, 2016, which claims the benefit of U.S. patentapplication Ser. No. 14/515,047, filed Oct. 15, 2014, which claims thebenefit of U.S. patent application Ser. No. 13/934,960, filed Jul. 3,2013, which claims the benefit of U.S. patent application Ser. No.12/562,621, filed Sep. 18, 2009, which claims the benefit of priority ofU.S. provisional application No. 61/097,896, filed Sep. 18, 2008, thedisclosures of which are hereby incorporated by reference as if writtenherein in their entireties.

Disclosed herein are new benzoquinoline compounds, pharmaceuticalcompositions made thereof, and methods to inhibit vesicular monoaminetransporter 2 (VMAT2) activity in a subject are also provided for, forthe treatment of chronic hyperkinetic movement disorders.

Tetrabenazine (Nitoman, Xenazine, Ro 1-9569),1,3,4,6,7,11b-Hexahydro-9,10-dimethoxy-3-(2-methylpropyl)-2H-benzo[a]quinoline,is a vesicular monoamine transporter 2 (VMAT2) inhibitor. Tetrabenazineis commonly prescribed for the treatment of Huntington's disease (Savaniet al., Neurology 2007, 68(10), 797; and Kenney et al., Expert Review ofNeurotherapeutics 2006, 6(1), 7-17).

In vivo, tetrabenazine is rapidly and extensively metabolized to itsreduced form,3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ol,which then binds specifically toVMAT2 (Zhang et al., AAPS Journal, 2006,8(4), E682-692). Additional metabolic pathways involve O-demethylationof the methoxy groups, as well as hydroxylation of the isobutyl group(Schwartz et al., Biochem. Pharmacol., 1966, 15, 645-655). Adverseeffects associated with the administration of tetrabenazine includeneuroleptic malignant syndrome, drowsiness, fatigue, nervousness,anxiety, insomnia, agitation, confusion, orthostatic hypotension,nausea, dizziness, depression, and Parkinsonism.

Deuterium Kinetic Isotope Effect

In order to eliminate foreign substances such as therapeutic agents, theanimal body expresses various enzymes, such as the cytochrome P₄₅₀enzymes (CYPs), esterases, proteases, reductases, dehydrogenases, andmonoamine oxidases, to react with and convert these foreign substancesto more polar intermediates or metabolites for renal excretion. Suchmetabolic reactions frequently involve the oxidation of acarbon-hydrogen (C—H) bond to either a carbon-oxygen (C—O) or acarbon-carbon (C—C) π-bond. The resultant metabolites may be stable orunstable under physiological conditions, and can have substantiallydifferent pharmacokinetic, pharmacodynamic, and acute and long-termtoxicity profiles relative to the parent compounds. For most drugs, suchoxidations are generally rapid and ultimately lead to administration ofmultiple or high daily doses.

The relationship between the activation energy and the rate of reactionmay be quantified by the Arrhenius equation, k=Ae^(−Eact/RT). TheArrhenius equation states that, at a given temperature, the rate of achemical reaction depends exponentially on the activation energy(E_(act)).

The transition state in a reaction is a short lived state along thereaction pathway during which the original bonds have stretched to theirlimit. By definition, the activation energy E_(act) for a reaction isthe energy required to reach the transition state of that reaction. Oncethe transition state is reached, the molecules can either revert to theoriginal reactants, or form new bonds giving rise to reaction products.A catalyst facilitates a reaction process by lowering the activationenergy leading to a transition state. Enzymes are examples of biologicalcatalysts.

Carbon-hydrogen bond strength is directly proportional to the absolutevalue of the ground-state vibrational energy of the bond. Thisvibrational energy depends on the mass of the atoms that form the bond,and increases as the mass of one or both of the atoms making the bondincreases. Since deuterium (D) has twice the mass of protium (¹H), a C-Dbond is stronger than the corresponding C—¹H bond. If a C—¹H bond isbroken during a rate-determining step in a chemical reaction (i.e. thestep with the highest transition state energy), then substituting adeuterium for that protium will cause a decrease in the reaction rate.This phenomenon is known as the Deuterium Kinetic Isotope Effect (DKIE).The magnitude of the DKIE can be expressed as the ratio between therates of a given reaction in which a C—¹H bond is broken, and the samereaction where deuterium is substituted for protium. The DKIE can rangefrom about 1 (no isotope effect) to very large numbers, such as 50 ormore. Substitution of tritium for hydrogen results in yet a strongerbond than deuterium and gives numerically larger isotope effects

Deuterium (²H or D) is a stable and non-radioactive isotope of hydrogenwhich has approximately twice the mass of protium (¹H), the most commonisotope of hydrogen. Deuterium oxide (D₂O or “heavy water”) looks andtastes like H₂O, but has different physical properties.

When pure D₂O is given to rodents, it is readily absorbed. The quantityof deuterium required to induce toxicity is extremely high. When about0-15% of the body water has been replaced by D₂O animals are healthy butare unable to gain weight as fast as the control (untreated) group. Whenabout 15-20% of the body water has been replaced with D₂O, the animalsbecome excitable. When about 20-25% of the body water has been replacedwith D₂O, the animals become so excitable that they go into frequentconvulsions when stimulated. Skin lesions, ulcers on the paws andmuzzles, and necrosis of the tails appear. The animals also become veryaggressive. When about 30% of the body water has been replaced with DO,the animals refuse to eat and become comatose. Their body weight dropssharply and their metabolic rates drop far below normal, with deathoccurring at about 30 to about 35% replacement with D₂O. The effects arereversible unless more than thirty percent of the previous body weighthas been lost due to D₂O. Studies have also shown that the use of D₂Ocan delay the growth of cancer cells and enhance the cytotoxicity ofcertain antineoplastic agents.

Deuteration of pharmaceuticals to improve pharmacokinetics (PK),pharmacodynamics (PD), and toxicity profiles has been demonstratedpreviously with some classes of drugs. For example, the DKIE was used todecrease the hepatotoxicity of halothane, presumably by limiting theproduction of reactive species such as trifluoroacetyl chloride.However, this method may not be applicable to all drug classes. Forexample, deuterium incorporation can lead to metabolic switching.Metabolic switching occurs when xenogens, sequestered by Phase Ienzymes, bind transiently and re-bind in a variety of conformationsprior to the chemical reaction (e.g., oxidation). Metabolic switching isenabled by the relatively vast size of binding pockets in many Phase Ienzymes and the promiscuous nature of many metabolic reactions.Metabolic switching can lead to different proportions of knownmetabolites as well as altogether new metabolites. This new metabolicprofile may impart more or less toxicity. Such pitfalls are non-obviousand are not predictable a priori for any drug class.

Tetrabenazine is a VMAT2 inhibitor. The carbon-hydrogen bonds oftetrabenazine contain a naturally occurring distribution of hydrogenisotopes, namely ¹H or protium (about 99.9844%), ²H or deuterium (about0.0156%), and ³H or tritium (in the range between about 0.5 and 67tritium atoms per 10¹⁸ protium atoms). Increased levels of deuteriumincorporation may produce a detectable Deuterium Kinetic Isotope Effect(DKIE) that could affect the pharmacokinetic, pharmacologic and/ortoxicologic profiles of tetrabenazine in comparison with tetrabenazinehaving naturally occurring levels of deuterium.

Based on discoveries made in our laboratory, as well as considering theliterature, tetrabenazine is metabolized in humans at the isobutyl andmethoxy groups. The current approach has the potential to preventmetabolism at these sites. Other sites on the molecule may also undergotransformations leading to metabolites with as-yet-unknownpharmacology/toxicology. Limiting the production of these metaboliteshas the potential to decrease the danger of the administration of suchdrugs and may even allow increased dosage and/or increased efficacy. Allof these transformations can occur through polymorphically-expressedenzymes, exacerbating interpatient variability. Further, some disordersare best treated when the subject is medicated around the clock or foran extended period of time. For all of the foregoing reasons, a medicinewith a longer half-life may result in greater efficacy and cost savings.Various deuteration patterns can be used to (a) reduce or eliminateunwanted metabolites, (b) increase the half-life of the parent drug, (c)decrease the number of doses needed to achieve a desired effect, (d)decrease the amount of a dose needed to achieve a desired effect, (e)increase the formation of active metabolites, if any are formed, (f)decrease the production of deleterious metabolites in specific tissues,and/or (g) create a more effective drug and/or a safer drug forpolypharmacy, whether the polypharmacy be intentional or not. Thedeuteration approach has the strong potential to slow the metabolism oftetrabenazine and attenuate interpatient variability.

Novel compounds and pharmaceutical compositions, certain of which havebeen found to inhibit VMAT2 have been discovered, together with methodsof synthesizing and using the compounds, including methods for thetreatment of VMAT2-mediated disorders in a patient by administering thecompounds as disclosed herein.

In certain embodiments of the present invention, compounds havestructural Formula I:

or a salt, solvate, or prodrug thereof, wherein:

R₁-R₂₇ are independently selected from the group consisting of hydrogenand deuterium; and

at least one of R₁-R27 is deuterium.

In certain embodiments, Formula I can include a single enantiomer, amixture of the (+)-enantiomer and the (−)-enantiomer, a mixture of about90% or more by weight of the (−)-enantiomer and about 10% or less byweight of the (+)-enantiomer, a mixture of about 90% or more by weightof the (+)-enantiomer and about 10% or less by weight of the(−)-enantiomer, an individual diastereomer, or a mixture ofdiastereomers thereof.

Certain compounds disclosed herein may possess useful VMAT2 inhibitingactivity, and may be used in the treatment or prophylaxis of a disorderin which VMAT2 plays an active role. Thus, certain embodiments alsoprovide pharmaceutical compositions comprising one or more compoundsdisclosed herein together with a pharmaceutically acceptable carrier, aswell as methods of making and using the compounds and compositions.Certain embodiments provide methods for inhibiting VMAT2. Otherembodiments provide methods for treating a VMAT2-mediated disorder in apatient in need of such treatment, comprising administering to saidpatient a therapeutically effective amount of a compound or compositionaccording to the present invention. Also provided is the use of certaincompounds disclosed herein for use in the manufacture of a medicamentfor the prevention or treatment of a disorder ameliorated by theinhibition of VMAT2.

The compounds as disclosed herein may also contain less prevalentisotopes for other elements, including, but not limited to, ¹³C or ¹⁴Cfor carbon, ³³S, ³⁴S, or ³⁶S for sulfur, ¹⁵N for nitrogen, and ¹⁷O or¹⁸O for oxygen.

In certain embodiments, the compound disclosed herein may expose apatient to a maximum of about 0.000005% D₂O or about 0.00001% DHO,assuming that all of the C-D bonds in the compound as disclosed hereinare metabolized and released as D₂O or DHO. In certain embodiments, thelevels of D₂O shown to cause toxicity in animals is much greater thaneven the maximum limit of exposure caused by administration of thedeuterium enriched compound as disclosed herein. Thus, in certainembodiments, the deuterium-enriched compound disclosed herein should notcause any additional toxicity due to the formation of D₂O or DHO upondrug metabolism.

In certain embodiments, the deuterated compounds disclosed hereinmaintain the beneficial aspects of the corresponding non-isotopicallyenriched molecules while substantially increasing the maximum tolerateddose, decreasing toxicity, increasing the half-life (T_(1/2)), loweringthe maximum plasma concentration (C_(max)) of the minimum efficaciousdose (MED), lowering the efficacious dose and thus decreasing thenon-mechanism-related toxicity, and/or lowering the probability ofdrug-drug interactions.

All publications and references cited herein are expressly incorporatedherein by reference in their entirety. However, with respect to anysimilar or identical terms found in both the incorporated publicationsor references and those explicitly put forth or defined in thisdocument, then those terms definitions or meanings explicitly put forthin this document shall control in all respects.

As used herein, the terms below have the meanings indicated.

The singular forms “a,” “an,” and “the” may refer to plural articlesunless specifically stated otherwise.

The term “about,” as used herein, is intended to qualify the numericalvalues which it modifies, denoting such a value as variable within amargin of error. When no particular margin of error, such as a standarddeviation to a mean value given in a chart or table of data, is recited,the term “about” should be understood to mean that range which wouldencompass the recited value and the range which would be included byrounding up or down to that figure as well, taking into accountsignificant figures.

When ranges of values are disclosed, and the notation “from n₁ . . . ton₂” or “n₁-n₂” is used, where n₁ and n₂ are the numbers, then unlessotherwise specified, this notation is intended to include the numbersthemselves and the range between them. This range may be integral orcontinuous between and including the end values.

The term “deuterium enrichment” refers to the percentage ofincorporation of deuterium at a given position in a molecule in theplace of hydrogen. For example, deuterium enrichment of 1% at a givenposition means that 1% of molecules in a given sample contain deuteriumat the specified position. Because the naturally occurring distributionof deuterium is about 0.0156%, deuterium enrichment at any position in acompound synthesized using non-enriched starting materials is about0.0156%. The deuterium enrichment can be determined using conventionalanalytical methods known to one of ordinary skill in the art, includingmass spectrometry and nuclear magnetic resonance spectroscopy.

The term “is/are deuterium,” when used to describe a given position in amolecule such as R₁-R₂₇ or the symbol “D”, when used to represent agiven position in a drawing of a molecular structure, means that thespecified position is enriched with deuterium above the naturallyoccurring distribution of deuterium. In one embodiment deuteriumenrichment is no less than about 1%, in another no less than about 5%,in another no less than about 10%, in another no less than about 20%, inanother no less than about 50%, in another no less than about 70%, inanother no less than about 80%, in another no less than about 90%, or inanother no less than about 98% of deuterium at the specified position.

The term “isotopic enrichment” refers to the percentage of incorporationof a less prevalent isotope of an element at a given position in amolecule in the place of the more prevalent isotope of the element.

The term “non-isotopically enriched” refers to a molecule in which thepercentages of the various isotopes are substantially the same as thenaturally occurring percentages.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as D-isomers and L-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Starting compounds ofparticular stereochemistry are either commercially available or can bemade and resolved by techniques known in the art. Additionally, thecompounds disclosed herein may exist as geometric isomers. The presentinvention includes all cis, trans, syn, anti, entgegen (E), and zusammen(Z) isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Additionally, the compounds disclosed herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The term “bond” refers to a covalent linkage between two atoms, or twomoieties when the atoms joined by the bond are considered to be part oflarger substructure. A bond may be single, double, or triple unlessotherwise specified. A dashed line between two atoms in a drawing of amolecule indicates that an additional bond may be present or absent atthat position.

The term “disorder” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disease”,“syndrome”, and “condition” (as in medical condition), in that allreflect an abnormal condition of the human or animal body or of one ofits parts that impairs normal functioning, is typically manifested bydistinguishing signs and symptoms.

The terms “treat,” “treating,” and “treatment” are meant to includealleviating or abrogating a disorder or one or more of the symptomsassociated with a disorder; or alleviating or eradicating the cause(s)of the disorder itself. As used herein, reference to “treatment”of adisorder is intended to include prevention. The terms “prevent,”“preventing,” and “prevention” refer to a method of delaying orprecluding the onset of a disorder; and/or its attendant symptoms,barring a subject from acquiring a disorder or reducing a subject's riskof acquiring a disorder.

The term “therapeutically effective amount” refers to the amount of acompound that, when administered, is sufficient to prevent developmentof, or alleviate to some extent, one or more of the symptoms of thedisorder being treated. The term “therapeutically effective amount” alsorefers to the amount of a compound that is sufficient to elicit thebiological or medical response of a cell, tissue, system, animal, orhuman that is being sought by a researcher, veterinarian, medicaldoctor, or clinician.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human, monkey, chimpanzee, gorilla, and the like),rodents (e.g., rats, mice, gerbils, hamsters, ferrets, and the like),lagomorphs, swine (e.g., pig, miniature pig), equine, canine, feline,and the like. The terms “subject” and “patient” are used interchangeablyherein in reference, for example, to a mammalian subject, such as ahuman patient.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a therapeutic disorder described in thepresent disclosure. Such administration encompasses co-administration ofthese therapeutic agents in a substantially simultaneous manner, such asin a single capsule having a fixed ratio of active ingredients or inmultiple, separate capsules for each active ingredient. In addition,such administration also encompasses use of each type of therapeuticagent in a sequential manner. In either case, the treatment regimen willprovide beneficial effects of the drug combination in treating thedisorders described herein.

The term “chronic hyperkinetic movement disorders” refers to disorderscharacterized by non-purposeful, repetitive, disordered motor acts,variously termed “compulsive”, “rhythmical”, or “stereotyped.” Inhumans, chronic hyperkinetic movement disorders can be psychogenic (e.g., tics), idiopathic (as in, e.g ., Tourette's syndrome and Parkinson'sDisease, genetic (as in, e.g ., the chorea characteristic ofHuntington's Disease), infectious (as in, e.g ., Sydenham's Chorea), or,as in tardive dyskinesia, drug-induced. Unless otherwise stated,“chronic hyperkinetic movement disorders” refers to and includes allpsychogenic, idiopathic, genetic, and drug-induced movement disorders.

The term “stereotyped” refers to a repeated behavior that appearsrepetitively with slight variation or, less commonly, as a complexseries of movements.

The term “VMAT2” refers to vesicular monoamine transporter 2, anintegral membrane protein that acts to transport monoamines—particularlyneurotransmitters such as dopamine, norepinephrine,serotonin, andhistamine—from cellular cytosol into synaptic vesicles.

The term “VMAT2-mediated disorder,” refers to a disorder that ischaracterized by abnormal VMAT2 activity. A VMAT2-mediated disorder maybe completely or partially mediated by modulating VMAT2. In particular,a VMAT2-mediated disorder is one in which inhibition of VMAT2 results insome effect on the underlying disorder e.g., administration of a VMAT2inhibitor results in some improvement in at least some of the patientsbeing treated.

The term “VMAT2 inhibitor”, “inhibit VMAT2”, or “inhibition of VMAT2”refers to the ability of a compound disclosed herein to alter thefunction of VMAT2. A VMAT2 inhibitor may block or reduce the activity ofVMAT2 by forming a reversible or irreversible covalent bond between theinhibitor and VMAT2 or through formation of a noncovalently boundcomplex. Such inhibition may be manifest only in particular cell typesor may be contingent on a particular biological event. The term “VMAT2inhibitor”, “inhibit VMAT2”, or “inhibition of VMAT2” also refers toaltering the function of VMAT2 by decreasing the probability that acomplex forms between a VMAT2 and a natural substrate

The term “therapeutically acceptable” refers to those compounds (orsalts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitablefor use in contact with the tissues of patients without excessivetoxicity, irritation, allergic response, immunogenecity, arecommensurate with a reasonable benefit/risk ratio, and are effective fortheir intended use.

The term “pharmaceutically acceptable carrier,” “pharmaceuticallyacceptable excipient,” “physiologically acceptable carrier,” or“physiologically acceptable excipient” refers to apharmaceutically-acceptable material, composition, or vehicle, such as aliquid or solid filler, diluent, excipient, solvent, or encapsulatingmaterial. Each component must be “pharmaceutically acceptable” in thesense of being compatible with the other ingredients of a pharmaceuticalformulation. It must also be suitable for use in contact with the tissueor organ of humans and animals without excessive toxicity, irritation,allergic response, immunogenecity, or other problems or complications,commensurate with a reasonable benefit/risk ratio. See, Remington: TheScience and Practice of Pharmacy, 21st Edition; Lippincott Williams &Wilkins: Philadelphia, Pa. , 2005; Handbook of PharmaceuticalExcipients, 5th Edition; Rowe et al., Eds., The Pharmaceutical Press andthe American Pharmaceutical Association: 2005; and Handbook ofPharmaceutical Additives, 3rd Edition; Ash and Ash Eds., GowerPublishing Company: 2007; Pharmaceutical Preformulation and Formulation,Gibson Ed., CRC Press LLC: Boca Raton, Fla., 2004).

The terms “active ingredient,” “active compound,” and “active substance”refer to a compound, which is administered, alone or in combination withone or more pharmaceutically acceptable excipients or carriers, to asubject for treating, preventing, or ameliorating one or more symptomsof a disorder.

The terms “drug,” “therapeutic agent,” and “chemotherapeutic agent”refer to a compound, or a pharmaceutical composition thereof, which isadministered to a subject for treating, preventing, or ameliorating oneor more symptoms of a disorder.

The term “release controlling excipient” refers to an excipient whoseprimary function is to modify the duration or place of release of theactive substance from a dosage form as compared with a conventionalimmediate release dosage form.

The term “nonrelease controlling excipient” refers to an excipient whoseprimary function do not include modifying the duration or place ofrelease of the active substance from a dosage form as compared with aconventional immediate release dosage form.

The term “prodrug” refers to a compound functional derivative of thecompound as disclosed herein and is readily convertible into the parentcompound in vivo. Prodrugs are often useful because, in some situations,they may be easier to administer than the parent compound. They may, forinstance, be bioavailable by oral administration whereas the parentcompound is not. The prodrug may also have enhanced solubility inpharmaceutical compositions over the parent compound. A prodrug may beconverted into the parent drug by various mechanisms, includingenzymatic processes and metabolic hydrolysis. See Harper, Progress inDrug Research 1962, 4, 221-294; Morozowich et al. in “Design ofBiopharmaceutical Properties through Prodrugs and Analogs,” Roche Ed.,APHA Acad. Pharm. Sci. 1977; “Bioreversible Carriers in Drug in DrugDesign, Theory and Application,” Roche Ed., APHA Acad. Pharm. Sci. 1987;“Design of Prodrugs,” Bundgaard, Elsevier, 1985; Wang et al., Curr.Pharm. Design 1999, 5, 265-287; Pauletti et al., Adv. Drug. DeliveryRev. 1997, 27, 235-256; Mizen et al., Pharm. Biotech. 1998, 11, 345-365;Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejad in“Transport Processes in Pharmaceutical Systems,” Amidon et al., Ed.,Marcell Dekker, 185-218, 2000; Balant et al., Eur. J. Drug Metab.Pharmacokinet. 1990, 15, 143-53; Balimane and Sinko, Adv. Drug DeliveryRev. 1999, 39, 183-209; Browne, Clin. Neuropharmacol. 1997, 20, 1-12;Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39; Bundgaard, Controlled DrugDelivery 1987, 17, 179-96; Bundgaard, Adv. Drug Delivery Rev.1992, 8,1-38; Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130;Fleisher et al., Methods Enzymol. 1985, 112, 360-381; Farquhar et al.,J. Pharm. Sci. 1983, 72, 324-325; Freeman et al., J. Chem. Soc., Chem.Commun. 1991, 875-877; Friis and Bundgaard, Eur. J. Pharm. Sci. 1996, 4,49-59; Gangwar et al., Des. Biopharm. Prop. Prodrugs Analogs, 1977,409-421; Nathwani and Wood, Drugs 1993, 45, 866-94; Sinhababu andThakker, Adv. Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs1985, 29, 455-73; Tan et al., Adv. Drug Delivery Rev. 1999, 39, 117-151;Taylor, Adv. Drug Delivery Rev. 1996, 19, 131-148; Valentino andBorchardt, Drug Discovery Today 1997, 2, 148-155; Wiebe and Knaus, Adv.Drug Delivery Rev. 1999, 39, 63-80; Waller et al., Br. J. Clin. Pharmac.1989, 28, 497-507.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The term “therapeutically acceptable salt,” as used herein,represents salts or zwitterionic forms of the compounds disclosed hereinwhich are therapeutically acceptable as defined herein. The salts can beprepared during the final isolation and purification of the compounds orseparately by reacting the appropriate compound with a suitable acid orbase.Therapeutically acceptable salts include acid and basic additionsalts. For a more complete discussion of the preparation and selectionof salts, refer to “Handbook of Pharmaceutical Salts, Properties, andUse,” Stah and Wermuth, Ed., (Wiley-VCH and VHCA, Zurich, 2002) andBerge et al., J. Pharm. Sci. 1977, 66, 1-19.

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, α-oxo-glutaric acid, glycolic acid,hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid,(+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid,maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, including, but not limited to, inorganic bases, such as magnesiumhydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, orsodium hydroxide; and organic bases, such as primary, secondary,tertiary, and quaternary, aliphatic and aromatic amines, includingL-arginine, benethamine, benzathine, choline, deanol, diethanolamine,diethylamine, dimethylamine, dipropylamine, diisopropylamine,2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine,isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine,morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine,piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine,pyridine, quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

While it may be possible for the compounds of the subject invention tobe administered as the raw chemical, it is also possible to present themas a pharmaceutical composition. Accordingly, provided herein arepharmaceutical compositions which comprise one or more of certaincompounds disclosed herein, or one or more pharmaceutically acceptablesalts, prodrugs, or solvates thereof, together with one or morepharmaceutically acceptable carriers thereof and optionally one or moreother therapeutic ingredients. Proper formulation is dependent upon theroute of administration chosen. Any of the well-known techniques,carriers, and excipients may be used as suitable and as understood inthe art; e.g., in Remington's Pharmaceutical Sciences. Thepharmaceutical compositions disclosed herein may be manufactured in anymanner known in the art, e.g., by means of conventional mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping or compression processes. The pharmaceuticalcompositions may also be formulated as a modified release dosage form,including delayed-, extended-, prolonged-, sustained-, pulsatile-,controlled-, accelerated- and fast-, targeted-, programmed-release, andgastric retention dosage forms. These dosage forms can be preparedaccording to conventional methods and techniques known to those skilledin the art (see, Remington: The Science and Practice of Pharmacy, supra;Modified-Release Drug Deliver Technology, Rathbone et al., Eds., Drugsand the Pharmaceutical Science, Marcel Dekker, Inc., New York, N.Y.,2002; Vol. 126).

The compositions include those suitable for oral, parenteral (includingsubcutaneous, intradermal, intramuscular, intravenous, intraarticular,and intramedullary), intraperitoneal, transmucosal, transdermal, rectaland topical (including dermal, buccal, sublingual and intraocular)administration although the most suitable route may depend upon forexample the condition and disorder of the recipient. The compositionsmay conveniently be presented in unit dosage form and may be prepared byany of the methods well known in the art of pharmacy. Typically, thesemethods include the step of bringing into association a compound of thesubject invention or a pharmaceutically salt, prodrug, or solvatethereof (“active ingredient”) with the carrier which constitutes one ormore accessory ingredients. In general, the compositions are prepared byuniformly and intimately bringing into association the active ingredientwith liquid carriers or finely divided solid carriers or both and then,if necessary, shaping the product into the desired formulation.

Formulations of the compounds disclosed herein suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. The active ingredient mayalso be presented as a bolus, electuary or paste.

Pharmaceutical preparations which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In addition, stabilizers may be added.Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. The formulations may be presentedin unit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in powder form or in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or sterile pyrogen-free water,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Formulations for parenteral administration include aqueous andnon-aqueous (oily) sterile injection solutions of the active compoundswhich may contain antioxidants, buffers, bacteriostats and solutes whichrender the formulation isotonic with the blood of the intendedrecipient; and aqueous and non-aqueous sterile suspensions which mayinclude suspending agents and thickening agents. Suitable lipophilicsolvents or vehicles include fatty oils such as sesame oil, or syntheticfatty acid esters, such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter, polyethylene glycol, or otherglycerides.

Certain compounds disclosed herein may be administered topically, thatis by non-systemic administration. This includes the application of acompound disclosed herein externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as gels, liniments, lotions, creams,ointments or pastes, and drops suitable for administration to the eye,ear or nose.

For administration by inhalation, compounds may be delivered from aninsufflator, nebulizer pressurized packs or other convenient means ofdelivering an aerosol spray. Pressurized packs may comprise a suitablepropellant such as dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Alternatively, foradministration by inhalation or insufflation, the compounds according tothe invention may take the form of a dry powder composition, for examplea powder mix of the compound and a suitable powder base such as lactoseor starch. The powder composition may be presented in unit dosage form,in for example, capsules, cartridges, gelatin or blister packs fromwhich the powder may be administered with the aid of an inhalator orinsufflator.

Preferred unit dosage formulations are those containing an effectivedose, as herein below recited, or an appropriate fraction thereof, ofthe active ingredient.

Compounds may be administered orally or via injection at a dose of from0.1 to 500 mg/kg per day. The dose range for adult humans is generallyfrom 5 mg to 2 g/day. Tablets or other forms of presentation provided indiscrete units may conveniently contain an amount of one or morecompounds which is effective at such dosage or as a multiple of thesame, for instance, units containing 5 mg to 500 mg, usually around 10mg to 200 mg.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration.

The compounds can be administered in various modes, e.g. orally,topically, or by injection. The precise amount of compound administeredto a patient will be the responsibility of the attendant physician. Thespecific dose level for any particular patient will depend upon avariety of factors including the activity of the specific compoundemployed, the age, body weight, general health, sex, diets, time ofadministration, route of administration, rate of excretion, drugcombination, the precise disorder being treated, and the severity of thedisorder being treated. Also, the route of administration may varydepending on the disorder and its severity.

In the case wherein the patient's condition does not improve, upon thedoctor's discretion the administration of the compounds may beadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisorder.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the compounds may be given continuouslyor temporarily suspended for a certain length of time (i.e., a “drugholiday”).

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved disorder is retained.Patients can, however, require intermittent treatment on a long-termbasis upon any recurrence of symptoms.

Disclosed herein are methods of treating a VMAT2-mediated disordercomprising administering to a subject having or suspected of having sucha disorder, a therapeutically effective amount of a compound asdisclosed herein or a pharmaceutically acceptable salt, solvate, orprodrug thereof.

VMAT2-mediated disorders, include, but are not limited to, chronichyperkinetic movment disorders, and/or any disorder which can lessened,alleviated, or prevented by administering a VMAT2 inhibitor. In certainembodiments, the chronic hyperkinetic movement disorder is Huntington'sdisease.

In certain embodiments, a method of treating a VMAT2-mediated disordercomprises administering to the subject a therapeutically effectiveamount of a compound as disclosed herein, or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof, so as to affect: (1)decreased inter-individual variation in plasma levels of the compound ora metabolite thereof; (2) increased average plasma levels of thecompound or decreased average plasma levels of at least one metaboliteof the compound per dosage unit; (3) decreased inhibition of, and/ormetabolism by at least one cytochrome P₄₅₀ or monoamine oxidase isoformin the subject; (4) decreased metabolism via at least onepolymorphically-expressed cytochrome P₄₅₀ isoform in the subject; (5) atleast one statistically-significantly improved disorder-control and/ordisorder-eradication endpoint; (6) an improved clinical effect duringthe treatment of the disorder, (7) prevention of recurrence, or delay ofdecline or appearance, of abnormal alimentary or hepatic parameters asthe primary clinical benefit, or (8) reduction or elimination ofdeleterious changes in any diagnostic hepatobiliary function endpoints,as compared to the corresponding non-isotopically enriched compound.

In certain embodiments, inter-individual variation in plasma levels ofthe compounds as disclosed herein, or metabolites thereof, is decreased;average plasma levels of the compound as disclosed herein are increased;average plasma levels of a metabolite of the compound as disclosedherein are decreased; inhibition of a cytochrome P₄₅₀ or monoamineoxidase isoform by a compound as disclosed herein is decreased; ormetabolism of the compound as disclosed herein by at least onepolymorphically-expressed cytochrome P₄₅₀ isoform is decreased; bygreater than about 5%, greater than about 10%, greater than about 20%,greater than about 30%, greater than about 40%, or by greater than about50% as compared to the corresponding non-isotopically enriched compound.

Plasma levels of the compound as disclosed herein, or metabolitesthereof, may be measured using the methods described by Li et al. RapidCommunications in Mass Spectrometry 2005, 19, 1943-1950; Jindal, et al.,Journal of Chromatography, Biomedical Applications 1989, 493(2), 392-7;Schwartz, et al., Biochemical Pharmacology 1966, 15(5), 645-55; Mehvar,et al., Drug Metabolism and Disposition 1987, 15(2), 250-5; Roberts etal., Journal of Chromatography, Biomedical Applications 1981, 226(1),175-82; and any references cited therein or any modifications madethereof.

Examples of cytochrome P₄₅₀ isoforms in a mammalian subject include, butare not limited to, CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6,CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2,CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11,CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1,CYP5A1, CYP7A1, CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2,CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39,CYP46, and CYP51.

Examples of monoamine oxidase isoforms in a mammalian subject include,but are not limited to, MAO_(A), and MAO_(B).

The inhibition of the cytochrome P₄₅₀ isoform is measured by the methodof Ko et al. (British Journal of Clinical Pharmacology, 2000, 49,343-351). The inhibition of the MAO_(A) isoform is measured by themethod of Weyler et al. (J. Biol Chem. 1985, 260, 13199-13207). Theinhibition of the MAO_(B) isoform is measured by the method of Uebelhacket al. (Pharmacopsychiatry, 1998, 31, 187-192).

Examples of polymorphically-expressed cytochrome P450 isoforms in amammalian subject include, but are not limited to, CYP2C8, CYP2C9,CYP2C19, and CYP2D6.

The metabolic activities of liver microsomes, cytochrome P₄₅₀ isoforms,and monoamine oxidase isoforms are measured by the methods describedherein.

Examples of improved disorder-control and/or disorder-eradicationendpoints, or improved clinical effects include, but are not limited to,change from baseline in the chorea score of the Unified Huntington'sDisease Rating Scale (UHDRS).

Examples of diagnostic hepatobiliary function endpoints include, but arenot limited to, alanine aminotransferase (“ALT”), serum glutamic-pyruvictransaminase (“SGPT”), aspartate aminotransferase (“AST” or “SGOT”),ALT/AST ratios, serum aldolase, alkaline phosphatase (“ALP”), ammonialevels, bilirubin, gamma-glutamyl transpeptidase (“GGTP,” “γ-GTP,” or“GGT”), leucine aminopeptidase (“LAP”), liver biopsy, liverultrasonography, liver nuclear scan, 5′-nucleotidase, and blood protein.Hepatobiliary endpoints are compared to the stated normal levels asgiven in “Diagnostic and Laboratory Test Reference”, 4^(th) edition,Mosby, 1999. These assays are run by accredited laboratories accordingto standard protocol.

Besides being useful for human treatment, certain compounds andformulations disclosed herein may also be useful for veterinarytreatment of companion animals, exotic animals and farm animals,including mammals, rodents, and the like. More preferred animals includehorses, dogs, and cats.

Combination Therapy

The compounds disclosed herein may also be combined or used incombination with other agents useful in the treatment of VMAT2-mediateddisorders. Or, by way of example only, the therapeutic effectiveness ofone of the compounds described herein may be enhanced by administrationof an adjuvant (i.e., by itself the adjuvant may only have minimaltherapeutic benefit, but in combination with another therapeutic agent,the overall therapeutic benefit to the patient is enhanced).

Such other agents, adjuvants, or drugs, may be administered, by a routeand in an amount commonly used therefor, simultaneously or sequentiallywith a compound as disclosed herein. When a compound as disclosed hereinis used contemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compounddisclosed herein may be utilized, but is not required.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more anti-psychotics, including, but not limited to,chlorpromazine, levomepromazine, promazine, acepromazine,triflupromazine, cyamemazine, chlorproethazine, dixyrazine,fluphenazine, perphenazine, prochlorperazine, thiopropazate,trifluoperazine, acetophenazine, thioproperazine, butaperazine,perazine, periciazine, thioridazine, mesoridazine, pipotiazine,haloperidol, trifluperidol, melperone, moperone, pipamperone,bromperidol, benperidol, droperidol, fluanisone, oxypertine, molindone,sertindole, ziprasidone, flupentixol, clopenthixol, chlorprothixene,thiothixene, zuclopenthixol, fluspirilene, pimozide, penfluridol,loxapine, clozapine, olanzapine, quetiapine, tetrabenazine, sulpiride,sultopride, tiapride, remoxipride, amisulpride, veralipride,levosulpiride, lithium, prothipendyl, risperidone, clotiapine,mosapramine, zotepine, pripiprazole, and paliperidone.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more benzodiazepines (“minor tranquilizers”), including, butnot limited to alprazolam, adinazolam, bromazepam, camazepam, clobazam,clonazepam, clotiazepam, cloxazolam, diazepam, ethyl loflazepate,estizolam, fludiazepam, flunitrazepam, halazepam, ketazolam, lorazepam,medazepam, dazolam, nitrazepam, nordazepam, oxazepam, potassiumclorazepate, pinazepam, prazepam, tofisopam, triazolam, temazepam, andchlordiazepoxide.

In certain embodiments, the compounds disclosed herein can be combinedwith olanzapine or pimozide.

The compounds disclosed herein can also be administered in combinationwith other classes of compounds, including, but not limited to,anti-retroviral agents; CYP3A inhibitors; CYP3A inducers; proteaseinhibitors; adrenergic agonists; anti-cholinergics; mast cellstabilizers; xanthines; leukotriene antagonists; glucocorticoidstreatments; local or general anesthetics; non-steroidalanti-inflammatory agents (NSAIDs), such as naproxen; antibacterialagents, such as amoxicillin; cholesteryl ester transfer protein (CETP)inhibitors, such as anacetrapib; anti-fungal agents, such asisoconazole; sepsis treatments, such as drotrecogin-α; steroidals, suchas hydrocortisone; local or general anesthetics, such as ketamine;norepinephrine reuptake inhibitors (NRIs) such as atomoxetine; dopaminereuptake inhibitors (DARIs), such as methylphenidate;serotonin-norepinephrine reuptake inhibitors (SNRIs), such asmilnacipran; sedatives, such as diazepham; norepinephrine-dopaminereuptake inhibitor (NDRIs), such as bupropion;serotonin-norepinephrine-dopamine-reuptake-inhibitors (SNDRIs), such asvenlafaxine; monoamine oxidase inhibitors, such as selegiline;hypothalamic phospholipids; endothelin converting enzyme (ECE)inhibitors, such as phosphoramidon; opioids, such as tramadol;thromboxane receptor antagonists, such as ifetroban; potassium channelopeners; thrombin inhibitors, such as hirudin; hypothalamicphospholipids; growth factor inhibitors, such as modulators of PDGFactivity; platelet activating factor (PAF) antagonists; anti-plateletagents, such as GPIIb/IIIa blockers (e.g., abdximab, eptifibatide, andtirofiban), P2Y(AC) antagonists (e.g., clopidogrel, ticlopidine andCS-747), and aspirin; anticoagulants, such as warfarin; low molecularweight heparins, such as enoxaparin; Factor VIIa Inhibitors and FactorXa Inhibitors; renin inhibitors; neutral endopeptidase (NEP) inhibitors;vasopepsidase inhibitors (dual NEP-ACE inhibitors), such as omapatrilatand gemopatrilat; HMG CoA reductase inhibitors, such as pravastatin,lovastatin, atorvastatin, simvastatin, NK-104 (a.k.a. itavastatin,nisvastatin, or nisbastatin), and ZD-4522 (also known as rosuvastatin,or atavastatin or visastatin); squalene synthetase inhibitors; fibrates;bile acid sequestrants, such as questran; niacin; anti-atheroscleroticagents, such as ACAT inhibitors; MTP Inhibitors; calcium channelblockers, such as amlodipine besylate; potassium channel activators;alpha-muscarinic agents; beta-muscarinic agents, such as carvedilol andmetoprolol; antiarrhythmic agents; diuretics, such as chlorothlazide,hydrochiorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorothiazide, trichioromethiazide,polythiazide, benzothlazide, ethacrynic acid, tricrynafen,chlorthalidone, furosenilde, musolimine, bumetanide, triamterene,amiloride, and spironolactone; thrombolytic agents, such as tissueplasminogen activator (tPA), recombinant tPA, streptokinase, urokinase,prourokinase, and anisoylated plasminogen streptokinase activatorcomplex (APSAC); anti-diabetic agents, such as biguanides (e.g.metformin), glucosidase inhibitors (e.g., acarbose), insulins,meglitinides (e.g., repaglinide), sulfonylureas (e.g., glimepiride,glyburide, and glipizide), thiozolidinediones (e.g. troglitazone,rosiglitazone and pioglitazone), and PPAR-gamma agonists;mineralocorticoid receptor antagonists, such as spironolactone andeplerenone; growth hormone secretagogues; aP2 inhibitors;phosphodiesterase inhibitors, such as PDE III inhibitors (e.g.,cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil,vardenafil); protein tyrosine kinase inhibitors; antiinflammatories;antiproliferatives, such as methotrexate, FK506 (tacrolimus, Prograf),mycophenolate mofetil; chemotherapeutic agents; immunosuppressants;anticancer agents and cytotoxic agents (e.g., alkylating agents, such asnitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimines, andtriazenes); antimetabolites, such as folate antagonists, purineanalogues, and pyrridine analogues; antibiotics, such as anthracyclines,bleomycins, mitomycin, dactinomycin, and plicamycin; enzymes, such asL-asparaginase; farnesyl-protein transferase inhibitors; hormonalagents, such as glucocorticoids (e.g., cortisone),estrogens/antiestrogens, androgens/antiandrogens, progestins, andluteinizing hormone-releasing hormone anatagonists, and octreotideacetate; microtubule-disruptor agents, such as ecteinascidins;microtubule-stablizing agents, such as pacitaxel, docetaxel, andepothilones A-F; plant-derived products, such as vinca alkaloids,epipodophyllotoxins, and taxanes; and topoisomerase inhibitors;prenyl-protein transferase inhibitors; and cyclosporins; steroids, suchas prednisone and dexamethasone; cytotoxic drugs, such as azathiprineand cyclophosphamide; TNF-alpha inhibitors, such as tenidap; anti-TNFantibodies or soluble TNF receptor, such as etanercept, rapamycin, andleflunimide; and cyclooxygenase-2 (COX-2) inhibitors, such as celecoxiband rofecoxib; and miscellaneous agents such as, hydroxyurea,procarbazine, mitotane, hexamethylmelamine, gold compounds, platinumcoordination complexes, such as cisplatin, satraplatin, and carboplatin.

Thus, in another aspect, certain embodiments provide methods fortreating VMAT2-mediated disorders in a subject in need of such treatmentcomprising administering to said subject an amount of a compounddisclosed herein effective to reduce or prevent said disorder in thesubject, in combination with at least one additional agent for thetreatment of said disorder. In a related aspect, certain embodimentsprovide therapeutic compositions comprising at least one compounddisclosed herein in combination with one or more additional agents forthe treatment of VMAT2-mediated disorders.

General Synthetic Methods for Preparing Compounds

Isotopic hydrogen can be introduced into a compound as disclosed hereinby synthetic techniques that employ deuterated reagents, wherebyincorporation rates are pre-determined; and/or by exchange techniques,wherein incorporation rates are determined by equilibrium conditions,and may be highly variable depending on the reaction conditions.Synthetic techniques, where tritium or deuterium is directly andspecifically inserted by tritiated or deuterated reagents of knownisotopic content, may yield high tritium or deuterium abundance, but canbe limited by the chemistry required. Exchange techniques, on the otherhand, may yield lower tritium or deuterium incorporation, often with theisotope being distributed over many sites on the molecule.

The compounds as disclosed herein can be prepared by methods known toone of skill in the art and routine modifications thereof, and/orfollowing procedures similar to those described in the Example sectionherein and routine modifications thereof, and/or procedures found inDaSilva et al., Appl. Radiat. Isot., 1993, 44(4), 673-676; Popp et al.,J. Pharm. Sci., 1978, 67(6), 871-873; Ivanov et al., Heterocycles 2001,55(8), 1569-1572; U.S. Pat. No. 2,830,993; U.S. Pat. No. 3,045,021; WO2007/130365; WO 2008/058261, which are hereby incorporated in theirentirety, and references cited therein and routine modificationsthereof. Compounds as disclosed herein can also be prepared as shown inany of the following schemes and routine modifications thereof.

The following schemes can be used to practice the present invention. Anyposition shown as hydrogen may be optionally substituted with deuterium.

Compound 1 is reacted with compound 2 in an appropriate solvent, such asnitromethane, in the presence of an appropriate acid, such as ammoniumacetate, at an elevated temperature to give compound 3. Compound 3 isreacted with compound 4 in the presence of an appropriate base, such aspotassium carbonate, in an appropriate solvent, such asN,N-dimethylformamide, at an elevated temperature to afford compound 5.Compound 5 is reacted with an appropriate reducing reagent, such aslithium aluminum hydride, in an appropriate solvent, such astetrahyrdofuran, at an elevated temperature to give compound 6. Compound6 is reacted with compound 7 in the presence of an appropriate acid,such as trifluoroacetic acid, in an appropriate solvent, such as aceticacid, at an elevated temperature to give compound 8. Compound 9 isreacted with compound 10 and compound 11, in an appropriate solvent,such as methanol, at an elevated temperature to afford compound 12.Compound 12 is reacted with compound 13 in an appropriate solvent, suchas ethyl acetate, to give compound 14. Compound 14 is reacted withcompound 8 in an appropriate solvent, such as ethanol, at an elevatedtemperature to give compound 15 of Formula I.

Deuterium can be incorporated to different positions synthetically,according to the synthetic procedures as shown in Scheme I, by usingappropriate deuterated intermediates. For example, to introducedeuterium at one or more positions of R₁-R₆, compound 4 with thecorresponding deuterium substitutions can be used. To introducedeuterium at one or more positions of R₇-R₉, compound 1 with thecorresponding deuterium substitutions can be used. To introducedeuterium at one or more positions of R₁₀ and R₁₂, lithium aluminumdeuteride can be used. To introduce deuterium at R_(11,) compound 2 withthe corresponding deuterium substitution can be used. To introducedeuterium at one or more positions of R₁₃-R_(14,) compound 10 with thecorresponding deuterium substitutions can be used. To introducedeuterium at R₁₅, compound 7 with the corresponding deuteriumsubstitution can be used. To introduce deuterium at one or morepositions of R₁₆-R_(27,) compound 9 with the corresponding deuteriumsubstitutions can be used.

Compound 8 is reacted with compound 16 in an appropriate solvent, suchas water, in the presence of an appropriate base, such as sodiumhydroxide, to yield a compound 15 of Formula I.

Deuterium can be incorporated to different positions synthetically,according to the synthetic procedures as shown in Scheme II, by usingappropriate deuterated intermediates. For example, to introducedeuterium at one or more positions of R₁-R₁₂ and R₁₅, compound 8 withthe corresponding deuterium substitutions can be used. To introducedeuterium at one or more positions of R₁₃-R₁₄ and R₁₆-R_(17,) andR₁₉-R_(27,) compound 16 with the corresponding deuterium substitutionscan be used. To introduce deuterium at R₁₈, deuterium oxide can be used.

Deuterium can also be incorporated to various positions, selectively ornon-selectively through a proton-deuterium exchange method known in theart.

The invention is further illustrated by the following examples. AllIUPAC names were generated using CambridgeSoft's ChemDraw 10.0.

EXAMPLE 13-Isobutyl-9,10-bis(methylamino)-3,4,6,7-tetrahydro-1H-pyrido[2,1-a]isoquinolin-2(11bH)-one(tetrabenazine)

Step 1

6,7-Dimethoxy-3,4-dihydroisoquinoline: Acetic acid/trifluoroacetic acid(100 mL) was added to a solution of 2-(3,4-dimethoxyphenyl)ethanamine(20 g, 110.38 mmol, 1.00 equiv) and hexamethylenetetramine (31 g, 221.11mmol, 2.00 equiv). The resulting solution was heated at reflux for about5 hours in an oil bath. After adding water (100 mL), the solution wasextracted with dichloromethane (3×200 mL), and the organic layers werecombined and dried over anhydrous sodium sulfate. Solids were removed byfiltration, and the resulting filtrate was concentrated in vacuo to givethe title product as a yellow oil (20 g, yield=95%). LC-MS :m/z=192(MH)⁺.

Step 2

3-((Dimethylamino)methyl)-5-methyl-hexan-2-one: Paraformaldehyde (5.5 g,183.33 mmol, 1.60 equiv) and dimethylamine hydrochloride (10 g, 122.70mmol, 1.00 equiv) were added to a solution of 5-methylhexan-2-one (50 g,437.83 mmol, 3.00 equiv) and methanol (30 mL). The resulting solutionwas heated at reflux for about 16 hours, and then the pH was adjusted toabout 8 with sodium hydroxide (10%). Standard extractive workup withether (3×100 mL) gave the title product as a yellow oil (12 g,yield=57%).

Step 3

(2-Acetyl-4-methyl-pentyl)-trimethyl-ammonium iodide: Iodomethane (20 g,140.94 mmol, 2.00 equiv) was added dropwise to a solution of3-((dimethylamino)methyl)-5-methylhexan-2-one (12 g, 70.05 mmol, 1.00equiv) and ethyl acetate (50 mL). The solution was stirred at ambienttemperature for about 16 hours., and the resulting preciptant wascollected by filtration to afford the title product (15 g, yield=68%).

Step 4

3-Isobutyl-9,10-dimethoxy-3,4,6,7-tetrahydro-1H-pyrido [2,1-alisoquinolin-2(11bH)-one: (2-Acetyl-4-methyl-pentyl)-trimethyl-ammoniumiodide (800 mg, 2.55 mmol, 1.00 equiv) was added to a solution of6,7-dimethoxy-3,4-dihydroisoquinoline (100 mg, 2.62 mmol, 1.00 equiv)and ethanol (10 mL). The resulting solution was heated at reflux forabout 5 hours, and then water (20 mL) was added. Following standardextractive workup with dichloromethane (3×50 mL), the crude residue waspurified by silica gel column chromatography (ethyl acetate/petroleumether (1:4)) to afford the title compound as a white solid (300 mg,yield=36%). 1H NMR (300 MHz, CDCl₃), δ6.63 (s, 1H), 6.55 (s, 1H), 3.89(s, 3H), 3.83 (s, 3H), 3.55 (s, 1H), 3.22-3.28 (m, 1H), 2.94-3.14 (m,4H), 2.31-2.65 (m, 4H), 1.73-1.81 (t, 1H , J=11.4), 1.33-1.39 (m, 1H),0.996-1.067 (t, 1H, J=10.5), 0.79-0.85 (m, 6H) LC-MS : m/z=318(MH)⁺.

EXAMPLE 23-Isobutyl-9,10-d₆-dimethoxy-3,4,6,7-tetrahydro-1H-pyrido[2,1-a]isoquinolin-2(11bH)-one(d₆-tetrabenazine)

Step 1

4-(2-Nitrovinyl)benzene-1,2-diol: Ammonium acetate (5.6 g, 72.73 mmol,1.00 equiv) was added to a solution of 3,4-dihydroxybenzaldehyde (10 g,72.41 mmol, 1.00 equiv) and nitromethane (50 mL). The resulting mixturewas heated at reflux for about 16 hours, and then concentrated in vacuo.The resulting residue was then purified by silica gel columnchromotagraphy (ethyl acetate/petroleum ether (1:5)) to give the titlecompound as a yellow oil (8 g, yield=61%).

Step 2

d₆-(E)-1,2-Dimethoxy-4-(2-nitrovinyl)benzene: d₃-Iodomethane (4.3 g, 31mmol, 1.10 equiv) was added dropwise to a solution of(E)-4-(2-nitrovinyl)benzene-1,2-diol (5 g, 28 mmol, 1.00 equiv),dimethylformamide (50 mL), and potassium carbonate (20 g, 140 mmol, 5.00equiv). The resulting suspension was stirred at about 50° C. for about16 hours. The suspension was filtered, the filter cake was washed withethyl acetate, and the washes were combined with the filtrate. Theresulting mixture was then extracted with ethyl acetate (200×3 mL) andwashed with brine. The mixture was dried over anhydrous sodium sulfateand concentrated in vacuo to give the title product as a yellow oil (5g, yield=77%).

Step 3

2-(3,4-d₆-Dimethoxyphenyl)ethanamine: At about 0° C., a solution ofd₆-(E)-1,2-dimethoxy-4-(2-nitrovinyl)benzene (8 g, 44.17 mmol, 1.00equiv) and tetrahydrofuran (20 mL) was added dropwise to a solution oflithium aluminum hydride (1.6 g, 42.11 mmol, 1.00 equiv) andtetrahydrofuran (30 mL). The resulting solution was heated at reflux forabout 16 hours, and then water (10 mL) was added. After removing solidsby filtation, the filtrate was dried over anhydrous magnesium sulfateand concentrated in vacuo to give the title product as a yellow solid (6g, yield=93%). LC-MS: m/z=188 (MH)⁺.

Step 4

6,7-d₆-Dimethoxy-3,4-dihydroisoquinoline: The procedure of Example 1,Step 1 was followed, but substituting2-(3,4-d₆-dimethoxyphenyl)ethanamine for2-(3,4-dimethoxyphenyl)ethanamine. The title product was isolated as ayellow oil (20 g, yield=95%).

Step 5

3-Isobutyl-9,10-d₆-dimethoxy-3,4,6,7-tetrahydro-1H-pyrido[2,1-a]isoquinolin-2(11bH)-one:The procedure of Example 1, Step 4 was followed but substituting6,7-d₆-dimethoxy-3,4-dihydroisoquinoline for6,7-dimethoxy-3,4-dihydroisoquinoline. The title product was isolated asa white solid (300 mg, yield=35%). 1H NMR (300 MHz, CDCl₃), δ 6.63 (s,1H), 6.55 (s, 1H), 3.55(s, 1H), 3.22-3.28 (m, 1H), 2.94-3.14 (m, 4H),2.31-2.65 (m, 4H), 1.73-1.81 (t, 1H , J=11.4), 1.33-1.39 (m, 1H),0.99-1.06 (t, 1H,=10.5), 0.79-0.85 (m, 6H) LC-MS: m/z=326 (MH)⁺.

The following compounds can generally be made using the methodsdescribed above. It is expected that these compounds when made will haveactivity similar to those described in the examples above.

Changes in the metabolic properties of the compounds disclosed herein ascompared to their non-isotopically enriched analogs can be shown usingthe following assays. Compounds listed above which have not yet beenmade and/or tested are predicted to have changed metabolic properties asshown by one or more of these assays as well.

Biological Activity Assays

In vitro Liver Microsomal Stability Assay

Liver microsomal stability assays are conducted at 1 mg per mL livermicrosome protein with an NADPH-generating system in 2% sodiumbicarbonate (2.2 mM NADPH, 25.6 mM glucose 6-phosphate, 6 units per mLglucose 6-phosphate dehydrogenase and 3.3 mM magnesium chloride). Testcompounds are prepared as solutions in 20% acetonitrile-water and addedto the assay mixture (final assay concentration 5 microgram per mL) andincubated at 37° C. Final concentration of acetonitrile in the assayshould be <1%. Aliquots (50 μL) are taken out at times 0, 15, 30, 45,and 60 minutes, and diluted with ice cold acetonitrile (200 μL) to stopthe reactions. Samples are centrifuged at 12,000 RPM for 10 minutes toprecipitate proteins. Supernatants are transferred to microcentrifugetubes and stored for LC/MS/MS analysis of the degradation half-life ofthe test compounds. It has thus been found that certaindeuterium-enriched compounds disclosed herein that have been tested inthis assay showed an increased degradation half-life as compared to thenon-isotopically enriched drug. In certain embodiments, the increase indegradation half-life is at least 5%.

In Vitro Metabolism Using Human Cytochrome P₄₅₀ Enzymes

The cytochrome P₄₅₀ enzymes are expressed from the corresponding humancDNA using a baculovirus expression system (BD Biosciences, San Jose,Calif.). A 0.25 milliliter reaction mixture containing 0.8 milligramsper milliliter protein, 1.3 millimolar NADP⁺, 3.3 millimolarglucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, 3.3millimolar magnesium chloride and 0.2 millimolar of a compound ofFormula I, the corresponding non-isotopically enriched compound orstandard or control in 100 millimolar potassium phosphate (pH 7.4) isincubated at 37° C. for 20 minutes. After incubation, the reaction isstopped by the addition of an appropriate solvent (e.g., acetonitrile,20% trichloroacetic acid, 94% acetonitrile/6% glacial acetic acid, 70%perchloric acid, 94% acetonitrile/6% glacial acetic acid) andcentrifuged (10,000 g) for 3 minutes. The supernatant is analyzed byHPLC/MS/MS.

Cytochrome P₄₅₀ Standard CYP1A2 Phenacetin CYP2A6 Coumarin CYP2B6[¹³C]-(S)-mephenytoin CYP2C8 Paclitaxel CYP2C9 Diclofenac CYP2C19[¹³C]-(S)-mephenytoin CYP2D6 (+/−)-Bufuralol CYP2E1 Chlorzoxazone CYP3A4Testosterone CYP4A [¹³C]-Lauric acid

Monoamine Oxidase A Inhibition and Oxidative Turnover

The procedure is carried out using the methods described by Weyler etal., Journal of Biological Chemistry 1985, 260, 13199-13207, which ishereby incorporated by reference in its entirety. Monoamine oxidase Aactivity is measured spectrophotometrically by monitoring the increasein absorbance at 314 nm on oxidation of kynuramine with formation of4-hydroxyquinoline. The measurements are carried out, at 30° C., in 50mM sodium phosphate buffer, pH 7.2, containing 0.2% Triton X-100(monoamine oxidase assay buffer), plus 1 mM kynuramine, and the desiredamount of enzyme in 1 mL total volume.

Monooamine Oxidase B Inhibition and Oxidative Turnover

The procedure is carried out as described in Uebelhack et al.,Pharmacopsychiatry 1998, 31(5), 187-192, which is hereby incorporated byreference in its entirety.

Determination of Tetrabenazine and an Active Metabolite by HPLC

The procedure is carried out as described in Roberts et al., Journal ofChromatography, Biomedical Applications 1981, 226(1), 175-82, which ishereby incorporated by reference in its entirety.

Pharmacokinetic Assays of Tetrabenazine and its Major Metabolite in Manand Rat

The procedure is carried out as described in Mehvar, et al., DrugMetabolism and Disposition 1987, 15(2), 250-5, which is herebyincorporated by reference in its entirety.

Detecting Tetrabenazine Metabolites in Animals and Man

The procedure is carried out as described in Schwartz, et al.,Biochemical Pharmacology 1966, 15(5), 645-55, which is herebyincorporated by reference in its entirety.

Mass Spectrometric Determination of Tetrabenazine

The procedure is carried out as described in Jindal, et al., Journal ofChromatography, Biomedical Applications 1989, 493(2), 392-7, which ishereby incorporated by reference in its entirety.

In Vitro Radioligand Binding Assay

The procedure is carried out as described in Scherman et al., Journal ofNeurochemistry 1988, 50(4), 1131-36, which is hereby incorporated byreference in its entirety.

In Vitro Radioligand Binding Assay

The procedure is carried out as described in Kilbourn et al., Synapse2002, 43(3), 188-194, which is hereby incorporated by reference in itsentirety.

In Vitro Radioligand Binding Assay

The procedure is carried out as described in Kilbourn et al., EuropeanJournal of Pharmacology 1997, 331(2-3), 161-68, which is herebyincorporated by reference in its entirety.

³H-Histamine Transport Assay

The procedure is carried out as described in Erickson et al., Journal ofMolecular Neuroscience 1995, 6(4), 277-87, which is hereby incorporatedby reference in its entirety.

From the foregoing description, one skilled in the art can ascertain theessential characteristics of this invention, and without departing fromthe spirit and scope thereof, can make various changes and modificationsof the invention to adapt it to various usages and conditions.

What is claimed is: 1-15. (canceled)
 16. A method of treatment of aVMAT2-mediated disorder comprising the administration to a subject of atherapeutically effective amount of a compound having the structuralformula


17. (canceled)
 18. (canceled)
 19. The method as recited in claim 16further comprising the administration of an additional therapeuticagent.
 20. The method as recited in claim 19 wherein said additionaltherapeutic agent is selected from the group consisting of olanzapineand pimozide.
 21. The method as recited in claim 19 wherein saidadditional therapeutic agent is selected from the group consisting ofbenzodiazepines, and antipsychotics.
 22. The method as recited in claim21 wherein said benzodiazepine is selected from the group consisting ofalprazolam, adinazolam, bromazepam, camazepam, clobazam, clonazepam,clotiazepam, cloxazolam, diazepam, ethyl loflazepate, estizolam,fludiazepam, flunitrazepam, halazepam, ketazolam, lorazepam, medazepam,dazolam, nitrazepam, nordazepam, oxazepam, potassium clorazepate,pinazepam, prazepam, tofisopam, triazolam, temazepam, andchlordiazepoxide.
 23. The method as recited in claim 21 wherein saidantipsychotic is selected from the group consisting of chlorpromazine,levomepromazine, promazine, acepromazine, triflupromazine, cyamemazine,chlorproethazine, dixyrazine, fluphenazine, perphenazine,prochlorperazine, thiopropazate, trifluoperazine, acetophenazine,thioproperazine, butaperazine, perazine, periciazine, thioridazine,mesoridazine, pipotiazine, haloperidol, trifluperidol, melperone,moperone, pipamperone, bromperidol, benperidol, droperidol, fluanisone,oxypertine, molindone, sertindole, ziprasidone, flupentixol,clopenthixol, chlorprothixene, thiothixene, zuclopenthixol,fluspirilene, pimozide, penfluridol, loxapine, clozapine, olanzapine,quetiapine, tetrabenazine, sulpiride, sultopride, tiapride, remoxipride,amisulpride, veralipride, levosulpiride, lithium, prothipendyl,risperidone, clotiapine, mosapramine, zotepine, pripiprazole, andpaliperidone. 24-33. (canceled)